Dendritic Cells and Periodontal Disease

树突状细胞和牙周病

• 批准号: 8476210
• 负责人: DANA T GRAVES
• 金额: $ 38.4万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8476210
• 关键词: Address; Alveolar Bone Loss; Anti-Inflammatory Agents; Anti-inflammatory; Antibodies; Bacteria; Binding; Biological Assay; Bone Resorption; Breeding; Calvaria; Cells; DNA; DNA Binding; Data; Dendritic Cells; Disease Progression; Down-Regulation; Event; Exhibits; Flow Cytometry; Gene Expression; Gene Targeting; Generations; Genetic Transcription; Goals; Histologic; ITGAX gene; Immune response; Immunization; In Vitro; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Interferons; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-6; Link; Measurement; Measures; Mediating; Modeling; Monitor; Mus; Nuclear; Oral; Oral cavity; Osteoclasts; Pathogenesis; Periodontal Diseases; Periodontium; Play; Porphyromonas gingivalis; Process; Production; Promoter Regions; Regulatory Pathway; Reporting; Role; Scalp structure; Signal Transduction; Site; Spleen; TNF gene; Testing; Up-Regulation; base; bone; bone loss; chromatin immunoprecipitation; cytokine; in vivo; in vivo Model; interleukin-12 subunit p40; mutant; oral infection; osteoclastogenesis; pathogen; prevent; promoter; recombinase; research study; response; soft tissue; transcription factor

DESCRIPTION (provided by applicant): Dendritic cells (DC) may play a central role in the pathogenesis of periodontal disease by participating in events that link bacterial stimulation to periodontal bone loss. They may accomplish this by the production of factors that regulate the adaptive and innate immune responses. Furthermore, immature DCs have been reported to function as osteoclasts precursors. Preliminary Data examining DCs in vitro establish that the transcription factor FOXO1 is essential for LPS up-regulation of several cytokines, IL-1, TNF, IL-6, and IL-12 and down-regulation of the anti-inflammatory cytokine IL-10. Moreover, Akt plays an important role in this process by reducing FOXO1 nuclear localization and limiting inflammatory cytokine production in DC. Thus, we propose that FOXO1 is critical in upregulating an inflammatory response in DCs and that a hyper- inflammatory response is prevented by Akt. These findings serve as the basis for the current proposal in which we will test in vivo the hypothesis that the FOXO1-Akt axis regulates DC cytokine expression. Moreover, we will determine whether this regulatory pathway is essential for stimulating the adaptive immune response to the periodontal pathogen, P. gingivalis, and whether it plays a significant role in bacteria induced bone loss. To delete FOXO1 in DCs we will use the Cre-lox approach. We have already bred floxed FOXO1 mice with mice that express Cre recombinase under control of the CD11c promoter. DC isolated from the resulting experimental (CD11cCre+/FOXO1L/L) mice exhibit reduced cytokine expression stimulated by LPS compared to littermate control mice (CD11cCre-/FOXO1L/L). We will examine host-bacteria interactions in vivo by injecting P. gingivalis in the calvarial model. Because mice are naive to P. gingivalis we can examine the response when the adaptive immune response is not present and compare it to mice in which the adaptive immune response is activated by pre-immunization with P. gingivalis. By flow cytometry we will have a detailed analysis on the impact of FOXO1 deletion in generating a systemic and local adaptive immune response. The underlying calvarial bone will also be examined histologically to investigate the subsequent effect on osteoclastogenesis and bone resorption. The oral gavage model of applying P. gingivalis to the oral cavity will be studied in experimental (CD11cCre+/FOXO1L/L) and control mice (CD11cCre-/FOXO1L/L ) to determine whether FOXO1 deletion in DCs modulates the host response to P. gingivalis, osteoclastogenesis and periodontal disease progression. Experiments using the same two in vivo models will determine whether Akt is necessary to prevent a hyperinflammatory response in DC. These experiments will examine experimental (CD11cCre+/FOXO1L/L) and control (CD11cCre-/FOXO1L/L) mice. In Aim 3 the mechanisms by which the FOXO1-Akt axis regulates selected target gene expression (IL-1, TNF, IL-6, and IL-12) will be examined in vitro.

描述（由申请人提供）：树突状细胞（DC）可能通过参与将细菌刺激与牙周骨质流失联系起来的事件，在牙周病的发病机制中发挥核心作用。他们可以通过产生调节适应性和先天免疫反应的因子来实现这一点。此外，据报道，未成熟的 DC 具有破骨细胞前体的功能。体外检查 DC 的初步数据表明，转录因子 FOXO1 对于 LPS 上调多种细胞因子（IL-1、TNF、IL-6 和 IL-12）以及下调抗炎细胞因子 IL-10 至关重要。此外，Akt 通过减少 FOXO1 核定位并限制 DC 中炎症细胞因子的产生，在此过程中发挥着重要作用。因此，我们认为 FOXO1 对于上调 DC 中的炎症反应至关重要，并且 Akt 可以预防高炎症反应。这些发现作为当前提议的基础，我们将在体内测试 FOXO1-Akt 轴调节 DC 细胞因子表达的假设。此外，我们将确定该调节途径是否对于刺激对牙周病原体牙龈卟啉单胞菌的适应性免疫反应至关重要，以及它是否在细菌引起的骨质流失中发挥重要作用。为了删除 DC 中的 FOXO1，我们将使用 Cre-lox 方法。我们已经用在 CD11c 启动子控制下表达 Cre 重组酶的小鼠培育了 floxed FOXO1 小鼠。与同窝对照小鼠 (CD11cCre-/FOXO1L/L) 相比，从所得实验 (CD11cCre+/FOXO1L/L) 小鼠中分离出的 DC 表现出 LPS 刺激的细胞因子表达减少。我们将通过在颅骨模型中注射牙龈卟啉单胞菌来检查体内宿主-细菌的相互作用。因为小鼠对牙龈卟啉单胞菌是幼稚的，所以我们可以检查当适应性免疫反应不存在时的反应，并将其与通过牙龈卟啉单胞菌预免疫激活适应性免疫反应的小鼠进行比较。通过流式细胞术，我们将详细分析 FOXO1 缺失对产生全身和局部适应性免疫反应的影响。还将对下面的颅骨进行组织学检查，以研究对破骨细胞生成和骨吸收的后续影响。将在实验小鼠 (CD11cCre+/FOXO1L/L) 和对照小鼠 (CD11cCre-/FOXO1L/L ) 中研究将牙龈卟啉单胞菌应用于口腔的口腔强饲模型，以确定树突状细胞中 FOXO1 缺失是否调节宿主对牙龈卟啉单胞菌、破骨细胞生成和牙周病进展的反应。使用相同的两个体内模型进行的实验将确定 Akt 是否对于预防 DC 的高炎症反应是必要的。这些实验将检查实验小鼠 (CD11cCre+/FOXO1L/L) 和对照小鼠 (CD11cCre-/FOXO1L/L)。在目标 3 中，将在体外检查 FOXO1-Akt 轴调节选定靶基因表达（IL-1、TNF、IL-6 和 IL-12）的机制。